Methods of inhibiting uterine fibrosis

ABSTRACT

A method of inhibiting uterine fibrosis comprising administering to a human or other mammal in need of treatment an effective amount of a compound having the formula ##STR1## Wherein R 1  and R 3  are independently hydrogen, ##STR2## wherein Ar is optionally substituted phenyl; wherein R 2  is selected from the group consisting of pyrrolidino and piperidino, a pharmaceutically acceptable salt or solvate thereof.

BACKGROUND OF THE INVENTION

Uterine fibrosis is an old and ever present clinical problem which goesunder a variety of names, including uterine hypertrophy, uterinelieomyomata, myometrial hypertrophy, fibrosis uteri, and fibroticmetritis. Essentially, uterine fibrosis is a condition where there is aninappropriate deposition of fibroid tissue on the wall of the uterus.

This condition is a cause of dysmenorrhea and infertility in women. Theexact cause of this condition is poorly understood but evidence suggeststhat it is an inappropriate response of fibroid tissue to estrogen. Sucha condition has been produced in rabbits by daily administrations ofestrogen for 3 months. In guinea pigs, the condition has been producedby daily administration of estrogen for four months. Further, in rats,estrogen causes similar hypertrophy.

The most common treatment of uterine fibrosis involves surgicalprocedures both costly and sometimes a source of complications such asthe formation of abdominal adhesions and infections. In some patients,initial surgery is only a temporary treatment and the fibroids regrow.In those cases a hysterectomy is performed which effectively ends thefibroids but also the reproductive life of the patient. Also,gonadotropin releasing hormone antagonists may be administered, yettheir use is tempered by the fact they can lead to osteoporosis.

SUMMARY OF THE INVENTION

This invention provides methods for inhibiting uterine fibrosis,comprising administering to a human or other mammal in need of treatmentan effective amount of a compound of formula I ##STR3## Wherein R¹ andR³ are independently hydrogen, ##STR4## wherein Ar is optionallysubstituted phenyl;

R² is selected from the group consisting of pyrrolidino and piperidino;and pharmaceutically acceptable salts and solyates thereof.

DETAILED DESCRIPTION OF THE INVENTION

The current invention concerns the discovery that a select group of2-phenyl-3-aroylbenzothiophenes (benzothiophenes), those of formula I,are useful for inhibiting uterine fibrosis. The methods of treatmentprovided by this invention are practiced by administering to a human inneed of inhibition of uterine fibrosis, a dose of a compound of formulaI or a pharmaceutically acceptable salt or solvate thereof, that iseffective to inhibit uterine fibrosis. The term inhibit is defined toinclude its generally accepted meaning which includes prophylacticallytreating a human subject to incurring uterine fibrosis, and holding incheck and/or treating existing uterine fibrosis. As such, the presentmethod includes both medical therapeutic and/or prophylactic treatment,as appropriate.

Generally, the compound is formulated with common excipients, diluentsor carriers, and compressed into tablets, or formulated as elixirs orsolutions for convenient oral administration, or administered by theintramuscular or intravenous routes. The compounds can be administeredtransdermally, and may be formulated as sustained release dosage formsand the like.

The compounds used in the methods of the current invention can be madeaccording to established procedures, such as those detailed in U.S. Pat.Nos. 4,133,814, 4,418,068, and 4,380,635 all of which are incorporatedby reference herein. In general, the process starts with abenzo[b]thiophene having a 6-hydroxyl group and a 2-(4-hydroxyphenyl)group. The starting compound is protected, alkylated, and deprotected toform the formula I compounds. Examples of the preparation of suchcompounds are provided in the U.S. patents discussed above. Substitutedphenyl includes phenyl substituted once or twice with C₁ -C₆ alkyl, C₁-C₄ alkoxy, hydromy, nitro, chloro, fluoro, or tri(chloro orfluoro)methyl.

The compounds used in the methods of this invention formpharmaceutically acceptable acid and base addition salts with a widevariety of organic and inorganic acids and bases and include thephysiologically acceptable salts which are often used in pharmaceuticalchemistry. Such salts are also part of this invention. Typical inorganicacids used to form such salts include hydrochloric, hydrobromic,hydroiodic, nitric, sulfuric, phosphoric, hypophosphoric and the like.Salts derived from organic acids, such as aliphatic mono anddicarboxylic acids, phenyl substituted alkanoic acids, hydroxyalkanoicand hydroxyalkandioic acids, aromatic acids, aliphatic and aromaticsulfonic acids, may also be used. Such pharmaceutically acceptable saltsthus include acetate, phenylacetate, trifluoroacetate, acrylate,ascorbate, benzoate, chlorobenzoate, dinitrobenzoate, hydroxybenzoate,methoxybenzoate, methylbenzoate, o-acetoxybenzoate,naphthalene-2-benzoate, bromide, isobutyrate, phenylbutyrate,β-hydroxybutyrate, butyne-1,4-dioate, hexyne-1,4-dioate, caprate,caprylate, chloride, cinnamate, citrate, formate, fumarate, glycollate,heptanoate, hippurate, lactate, malate, maleate, hydroxymaleate,malonate, mandelate, mesylate, nicotinate, isonicotinate, nitrate,oxalate, phthalate, teraphthalate, phosphate, monohydrogenphosphate,dihydrogenphosphate, metaphosphate, pyrophosphate, propiolate,propionate, phenylpropionate, salicylate, sebacate, succinate, suberate,sulfate, bisulfate, pyrosulfate, sulfite, bisulfite, sulfonate,benzene-sulfonate, p-bromophenylsulfonate, chlorobenzenesulfonate,ethanesulfonate, 2-hydroxyethanesulfonate, methanesulfonate,naphthalene-1-sulfonate, naphthalene-2-sulfonate, p-toluenesulfonate,xylenesulfonate, tartarate, and the like. A preferable salt is thehydrochloride salt.

The pharmaceutically acceptable acid addition salts are typically formedby reacting a compound of formula I with an equimolar or excess amountof acid. The reactants are generally combined in a mutual solvent suchas diethyl ether or benzene. The salt normally precipitates out ofsolution within about one hour to 10 days and can be isolated byfiltration or the solvent can be stripped off by conventional means.

Bases commonly used for formation of salts include ammonium hydroxideand alkali and alkaline earth metal hydroxides and carbonates, as wellas aliphatic and aromatic amines, aliphatic diamines and hydroxyalkylamines. Bases especially useful in the preparation of additionsalts include ammonium hydroxide, potassium carbonate, sodiumbicarbonate, calcium hydroxide, methylamine, diethylamine, ethylenediamine, cyclohexylamine and ethanolamine.

The pharmaceutically acceptable salts generally have enhanced solubilitycharacteristics compared to the compound from which they are derived,and thus are often more amenable to formulation as liquids or emulsions.

Pharmaceutical formulations can be prepared by procedures known in theart. For example, the compounds can be formulated with commonexcipients, diluents, or carriers, and formed into tablets, capsules,suspensions, powders, and the like. Examples of excipients, diluents,and carriers that are suitable for such formulations include thefollowing: fillers and extenders such as starch, sugars, mannitol, andsilicic derivatives; binding agents such as carboxymethyl cellulose andother cellulose derivatives, alginates, gelatin, and polyvinylpyrrolidone; moisturizing agents such as glycerol; disintegrating agentssuch as agaragar, calcium carbonate, and sodium bicarbonate; agents forretarding dissolution such as paraffin; resorption accelerators such asquaternary ammonium compounds; surface active agents such as cetylalcohol, glycerol monostearate; adsorptive carriers such as kaolin andbentonire; and lubricants such as talc, calcium and magnesium stearate,and solid polyethyl glycols.

The compounds can also be formulated as elixirs or solutions forconvenient oral administration or as solutions appropriate forparenteral administration, for instance by intramuscular, subcutaneousor intravenous routes. Additionally, the compounds are well suited toformulation as sustained release dosage forms and the like. Theformulations can be so constituted that they release the activeingredient only or preferably in a particular part of the intestinaltract, possibly over a period of time. The coatings, envelopes, andprotective matrices may be made, for example, from polymeric substancesor waxes.

The particular dosage of a compound of formula I required to inhibituterine fibrosis, according to this invention will depend upon theseverity of the condition, the route of administration, and relatedfactors that will be decided by the attending physician. Generally,accepted and effective daily doses will be from about 0.1 to about 1000mg/day, and more typically from about 50 to about 200 mg/day. Suchdosages will be administered to a subject in need of treatment from onceto about three times each day, or more often as needed to effectivelyinhibit uterine fibrosis.

The invention also provides for a method of inhibiting uterine fibrosisby administering serially or concurrently a compound of the inventionand a gonadotropin releasing hormone antangonist.

It is usually preferred to administer a compound of formula I in theform of an acid addition salt, as is customary in the administration ofpharmaceuticals bearing a basic group, such as the piperidino ring. Forsuch purposes the following dosage forms are available.

FORMULATIONS

In the formulations which follow, "Active ingredient" means a compoundof formula I.

Formulation 1: Gelatin Capsules Hard gelatin capsules are prepared usingthe following:

    ______________________________________                                        Ingredient        Quantity (mg/capsule)                                       ______________________________________                                        Active ingredient 0.1-1000                                                    Starch, NF        0-650                                                       Starch flowable powder                                                                          0-650                                                       Silicone fluid 350 centistokes                                                                  0-15                                                        ______________________________________                                    

The ingredients are blended, passed through a No. 45 mesh U.S. sieve,and filled into hard gelatin capsules.

Examples of specific capsule formulations of the compound of formula 1wherein R² is piperidino, (raloxifene), that have been made includethose shown below:

Formulation 2: Raloxifene capsule

    ______________________________________                                        Ingredient        Quantity (mg/capsule)                                       ______________________________________                                        Raloxifene        1                                                           Starch, NF        112                                                         Starch flowable powder                                                                          225.3                                                       Silicone fluid 350 centistokes                                                                  1.7                                                         ______________________________________                                    

Formulation 3: Raloxifene capsule

    ______________________________________                                        Ingredient        Quantity (mg/capsule)                                       ______________________________________                                        Raloxifene        5                                                           Starch, NF        108                                                         Starch flowable powder                                                                          225.3                                                       Silicone fluid 350 centistokes                                                                  1.7                                                         ______________________________________                                    

Formulation 4: Raloxifene capsule

    ______________________________________                                        Ingredient        Quantity (mg/capsule)                                       ______________________________________                                        Raloxifene        10                                                          Starch, NF        103                                                         Starch flowable powder                                                                          225.3                                                       Silicone fluid 350 centistokes                                                                  1.7                                                         ______________________________________                                    

Formulation 5: Raloxifene capsule

    ______________________________________                                        Ingredient        Quantity (mg/capsule)                                       ______________________________________                                        Raloxifene        50                                                          Starch, NF        150                                                         Starch flowable powder                                                                          397                                                         Silicone fluid 350 centistokes                                                                  3.0                                                         ______________________________________                                    

The specific formulations above may be changed in compliance with thereasonable variations provided.

A tablet formulation is prepared using the ingredients below:

Formulation 6: Tablets

    ______________________________________                                        Ingredient      Quantity (mg/tablet)                                          ______________________________________                                        Active ingredient                                                                             0.1-1000                                                      Cellulose, micro-                                                                             0-650                                                         crystalline                                                                   Silicon dioxide, fumed                                                                        0-650                                                         Stearate acid   0-15                                                          ______________________________________                                    

The components are blended and compressed to form tablets.

Alternatively, tablets each containing 0.1-1000 mg of active ingredientare made up as follows:

Formulation 7: Tablets

    ______________________________________                                        Ingredient        Quantity (mg/tablet)                                        ______________________________________                                        Active ingredient 0.1-1000                                                    Starch            45                                                          Cellulose, microcrystalline                                                                     35                                                          Polyvinylpyrrolidone                                                                            4                                                           (as 10% solution in water)                                                    Sodium carboxymethyl cellulose                                                                  4.5                                                         Magnesium stearate                                                                              0.5                                                         Talc              1                                                           ______________________________________                                    

The active ingredient, starch, and cellulose are passed through a No. 45mesh U.S. sieve and mixed thoroughly. The solution ofpolyvinylpyrrolidone is mixed with the resultant powders which are thenpassed through a No. 14 mesh U.S. sieve. The granules so produced aredried at 50°-60° C. and passed through a No. 18 mesh U.S. sieve. Thesodium carboxymethyl starch, magnesium stearate, and talc, previouslypassed through a No. 60 U.S. sieve, are then added to the granuleswhich, after mixing, are compressed on a tablet machine to yieldtablets.

Suspensions each containing 0.1-1000 mg of medicament per 5 mL dose aremade as follows:

Formulation 8: Suspensions

    ______________________________________                                        Ingredient            Quantity (mg/5 ml)                                      ______________________________________                                        Active ingredient     0.1-1000  mg                                            Sodium carboxymethyl cellulose                                                                      50        mg                                            Syrup                 1.25      mg                                            Benzoic acid solution 0.10      mL                                            Flavor                q.v.                                                    Color                 q.v.                                                    Purified water to     5         mL                                            ______________________________________                                    

The medicament is passed through a No. 45 mesh U.S. sieve and mixed withthe sodium carboxymethyl cellulose and syrup to form a smooth paste. Thebenzoic acid solution, flavor, and color are diluted with some of thewater and added, with stirring. Sufficient water is then added toproduce the required volume.

Test 1

Between 3 and 20 women having uterine fibrosis are administered acompound of the invention. The amount of compound administered is 0.1 to1000 mg/day, and the period of administration is 3 months.

The women are observed during the period of administration and up to 3months after discontinuance of the compound for effects on uterinefibrosis.

Test 2

The same procedure is used as in Test 1, except the period ofadministration is 6 months.

Test 3

The same procedure is used as in Test 1, except the period ofadministration is 1 year.

Test 4

A. Induction of fibroid tumors in guinea pig.

Prolonged estrogen stimulation is used to induce leiomyomata in sexuallymature female guinea pigs. Animals are dosed with estradiol 3-5 timesper week by injection for 2-4 months or until tumors arise. Treatmentsconsisting of a compound of the invention or vehicle is administereddaily for 3-16 weeks and then animals are sacrificed and the uteriharvested and analyzed for tumor regression.

B. Implantation of human uterine fibroid tissue in nude mice.

Tissue from human leiomyomas are implanted into the peritoneal cavityand or uterine myometrium of sexually mature, castrated female nudemice. Exogenous estrogen are supplied to induce growth of the explantedtissue. In some cases the harvested tumor cells are cultured in vitroprior to implantation. Treatment consisting of a compound of theinvention or vehicle is supplied by gastric lavage on a daily basis for3-16 weeks and implants are removed and measured for growth orregression. At the time of sacrifice, the uteri is harvested to assessthe status of the organ.

Test 5

A. Tissue from human uterine fibroid tumors is harvested, and maintainedin vitro as primary nontransformed cultures. Surgical specimens arepushed through a sterile mesh or sieve or alternately teased apart fromsurrounding tissue to produce a single cell suspension. Cells aremaintained in media containing 10% serum and antibiotics. Rates ofgrowth in the presence and absence of estrogen are determined. Cells areassayed for their ability to produce complement component C3 and theirresponse to growth factors and growth hormone. In vitro cultures areassessed for their proliferative response following treatment withprogestins, GnRH, a compound of the invention and vehicle. Levels ofsteroid hormone receptors are assessed weekly to determine whetherimportant cell characteristics are maintained in vitro. Tissue from 5-25patients are utilized.

Activity in at least one of the above tests indicates the compounds ofthe invention are of potential in the treatment of uterine fibrosis.

We claim:
 1. A method of inhibiting uterine fibrosis comprisingadministering to a human or other mammal in need of treatment aneffective amount of a compound having the formula ##STR5## Wherein R¹and R³ are independently hydrogen, ##STR6## wherein Ar is optionallysubstituted phenyl; R² is selected from the group consisting ofpyrrolidino and piperidino; or a pharmaceutically acceptable salt orsolvate thereof.
 2. The method of claim 1 wherein said compound is thehydrochloride salt thereof.
 3. The method of claim 1 wherein saidcompound is ##STR7## or its hydrochloride salt.
 4. The method accordingto claim 1 wherein said human or other mammal is also administeredgonadotropin releasing hormone antagonist.